Inhibiting disulphide bonding in truncated tau297-391 results in enhanced self-assembly of tau into seed-competent assemblies.

Oakley, S.; Marshall, K. E.; Meisl, G. E.; Maina, M. B.; Milton, R.; Vorley, T.; Storey, J.; Harrington, C.; WISCHIK, C. M.; Xue, W.-F.; Copsey, A.; Serpell, L.

doi:10.1101/2025.02.05.636249

bioRxiv · 10.1101/2025.02.05.636249

Inhibiting disulphide bonding in truncated tau297-391 results in enhanced self-assembly of tau into seed-competent assemblies.

Oakley, S.·Marshall, K. E.·Meisl, G. E.·Maina, M. B.·Milton, R.·Vorley, T.·Storey, J.·Harrington, C.·WISCHIK, C. M.·Xue, W.-F.·Copsey, A.·Serpell, L.

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Abstract

Tau undergoes fibrillogenesis in a group of neurodegenerative diseases termed tauopathies. Each tauopathy is characterized by tau fibrils with disease-specific conformations, highlighting the complexity of tau self-assembly. This has led to debate surrounding the precise mechanisms that govern the self-assembly of tau in disease, especially the involvement of disulphide bonding (DSB) between cysteine residues. In this study, we use a truncated form of tau, dGAE, capable of forming filaments identical to those in disease. We reveal the impact of DSB in dGAE assembly and propagation by resolving the global mechanisms that dominate its assembly. We found evidence for surface-mediated secondary nucleation and fragmentation being active in dGAE assembly. The inhibition of DSB during dGAE assembly leads to an enhanced aggregation rate through a reduced lag phase, but with no effect on the global assembly mechanisms. We suggest this is due to the formation a dominant, seed-competent species in the absence of DSB that facilitates elongation and secondary nucleation resulting in enhanced assembly. In vitro seeding assays reveal the recruitment of endogenous tau in a cell model only when using dGAE species formed under conditions that inhibit DSB. Our results further support the use of the in vitro dGAE tau aggregation model for investigating the mechanism of tau assembly, the effect of varying conditions on tau assembly and how these conditions affect the resultant species. Further studies may utilise dGAE and its aggregates to investigate tau seeding, propagation and to highlight or test potential targets for therapies that reduce the spread of pathologic tau throughout the brain.

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Oakley, S., Marshall, K. E., Meisl, G. E., Maina, M. B., Milton, R., Vorley, T., Storey, J., Harrington, C., WISCHIK, C. M., Xue, W.-F., Copsey, A., Serpell, L.. 2025-02-05. Inhibiting disulphide bonding in truncated tau297-391 results in enhanced self-assembly of tau into seed-competent assemblies.. https://doi.org/10.1101/2025.02.05.636249

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